EP1117408A1 - Use of diltiazem for treating retinal pathologies - Google Patents

Use of diltiazem for treating retinal pathologies

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Publication number
EP1117408A1
EP1117408A1 EP99946279A EP99946279A EP1117408A1 EP 1117408 A1 EP1117408 A1 EP 1117408A1 EP 99946279 A EP99946279 A EP 99946279A EP 99946279 A EP99946279 A EP 99946279A EP 1117408 A1 EP1117408 A1 EP 1117408A1
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EP
European Patent Office
Prior art keywords
diltiazem
enantiomer
photoreceptors
pharmaceutically acceptable
cis
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
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EP99946279A
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German (de)
French (fr)
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EP1117408B9 (en
EP1117408B1 (en
Inventor
Serge Picaud
Maria Frasson
José SAHEL
Henri Dreyfus
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Universite Louis Pasteur Strasbourg I
Institut National de la Sante et de la Recherche Medicale INSERM
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Universite Louis Pasteur Strasbourg I
Institut National de la Sante et de la Recherche Medicale INSERM
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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A61K31/554—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole having at least one nitrogen and one sulfur as ring hetero atoms, e.g. clothiapine, diltiazem
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents

Definitions

  • the present invention relates to the use of calcium channel blocking compounds and / or channels activated by cyclic guanosine monophosphate 3 '5' (cGMP) in the context of the treatment of pathologies of the retina, and more particularly of diseases of the retina due to a degeneration of photoreceptors, in humans or animals, such as retinitis pigmentosa or other pathologies similarly affecting photoreceptors, in particular macular degeneration linked to age.
  • cGMP cyclic guanosine monophosphate 3 '5'
  • Pigmentary retinitis is a group of these degenerative photoreceptor diseases (Berson, 1996) leading to blindness. Numerous mutations affecting different rod proteins, and likely to be at the origin of this disease, have been highlighted. Among these mutations, mention may be made of those affecting the genes of proteins involved in the phototransduction cascade, such as rhodopsin, transducin, phosphodiesterase, rarrestine, or of structural proteins, such as peripherin.
  • the rd mouse (retinal degeneration) has been studied for more than 70 years as a model of pigmentary retinitis (Farber et al., 1994) because the process of retinal degeneration is similar to that observed in the pigment retina, the death of the rods of the retina being followed by an unexplained loss of cones in the retina.
  • the causal mutation was located in the gene coding for the ⁇ subunit of cGMP-phosphodiesterase (PDE)
  • PDE is activated during the phototransduction cascade by the ⁇ chain of transducin, itself activated by light-stimulated rhodopsin. Activated PDE hydrolyzes cGMP, thereby reducing the concentration of cGMP and therefore the number of open cGMP-dependent channels, the final consequence being
  • Murine PDE (Bennett et al., 1996). Transplantation of photoreceptors (Gouras et al., 1994, Silverman et al., 1989) has been described as making it possible to preserve the cone photoreceptors (Mohand-Said et al., 1997). The interpretation of this effect as a paracrine mechanism is correlated with the increase in photoreceptor survival observed in coculture with healthy photoreceptors (Mohand-Said et al., 1998) or after in vivo or in vitro application of factors trophics such as fibroblast or neuronal growth factors (LaVail et al., 1998).
  • the object of the present invention is precisely to provide pharmaceutical compositions which can be used in the treatment of diseases of the retina due to degeneration of photoreceptors in humans or animals. Indeed, the present invention follows from the demonstration by the present invention.
  • the calcium channel and / or cGMP-dependent mocking compounds make it possible not only to slow the degeneration of rods and cones in rd mice, but also to preserve the capacity of retinal cells to respond to light stimuli.
  • the subject of the present invention is the use of calcium channel blocking and / or cGMP-dependent compounds for the preparation of a medicament intended for the treatment of pathologies linked to the degeneration of photoreceptors of the retina, and more particularly of the retinitis pigmentosa, or pathologies similarly affecting photoreceptors, such as age-related macular degeneration.
  • blocking compounds of calcium channels and / or cGMP-dependent is meant any compound capable of reducing the ionic conductance of these channels.
  • a more particular subject of the invention is the use of diltiazem (D-cis-enantiomer), L-cis-enantiomer, their metabolites and their pharmaceutically acceptable salts, for the preparation of a medicament intended for the treatment of pathologies linked to the degeneration of the photoreceptors of the retina, in particular to the treatment of retinitis pigmentosa, or pathologies similarly affecting photoreceptors, such as age-related macular degeneration.
  • D-cis-enantiomer D-cis-enantiomer
  • L-cis-enantiomer L-cis-enantiomer
  • their metabolites and their pharmaceutically acceptable salts
  • the invention relates more particularly to the aforementioned use of the diltiazem of formula
  • the subject of the invention is also the above-mentioned use of the L-cis enantiomer of diltiazem, or the racemate of diltiazem, as well as their addition salts with pharmaceutically acceptable acids, in particular the hydrochloride of the cis (-) isomer of formula next :
  • the invention also relates to the abovementioned use of one of the metabolites of diltiazem, as well as their addition salts with pharmaceutically acceptable acids, corresponding to the following formulas:
  • the above-mentioned calcium channel blocking and / or cGMP-dependent compounds are used for the preparation of pharmaceutical compositions which are in a form which can be administered by any route, in particular by oral, intramuscular, intravenous, intraocular, or in the form of eye drops.
  • the pharmaceutical compositions of the invention comprise, in unit form, approximately 0.1 to approximately 100 mg of a calcium channel blocking compound and / or cGMP-dependent, as defined above, in combination with a vehicle pharmaceutically acceptable.
  • FIG. 1 illustrates the survival of the rods at the 25th day and at the 36th postnatal day in rd mice treated with diltiazem hydrochloride.
  • the injections of diltiazem hydrochloride started on the 9th postnatal day which corresponds to the period of appearance of the first signs of degeneration of the rods.
  • the doses of diltiazem hydrochloride (2.5 mg / ml in physiological solution) were gradually increased from 50 ⁇ l per day to 100 ⁇ l twice a day following the growth of the animal. Since the electroretinograms (ERG) were measured before the animals were sacrificed, the last injection was given 48 hours before the physiological measurement.
  • ERP electroretinograms
  • the rods were labeled with anti-rhodopsin antibodies (rho-4D2) (Hicks and Molday, 1986) and their number was estimated by stereology on the retinas laid flat using a random sampling procedure. (Mohand-Said et al., 1998). Only the right retinas were taken into account in order to obtain independent results. Treatment with diltiazem hydrochloride increased the survival of the rods at day 25 by 86%, and by 148% at day 36 in the treated animals compared to the control rd mice.
  • rho-4D2 anti-rhodopsin antibodies
  • FIG. 2 illustrates the survival of the cones on the 25th day and on the 36th postnatal day in rd mice treated with diltiazem hydrochloride.
  • the number of cones was estimated indirectly after staining of the nuclei with DAPI (4 ', 6-diamino-2-phenylindole) on sections of retina. To obtain this number, the number of immunostained rods was subtracted from the number of photoreceptor nuclei labeled with DAPI.
  • Treatment with diltiazem hydrochloride increased the survival of the cones at the 25th day by 109% and by 144% at the 36th day in the treated animals compared to the control rd mice.
  • FIG. 3 illustrates the fact that the survival of photoreceptors is accompanied by physiological improvements in rd mice treated with diltiazem hydrochloride.
  • This demonstration was carried out by measurement of ERG in the rd treated mice and the rd control mice.
  • the amplitudes of the a and b waves of the ERGs decrease regularly from the 12th postnatal day until extinction on the 24th postnatal day.
  • 4 animals out of 10 treated show ERG signals which can be measured in both eyes.
  • FIG. 3 effect of diltiazem hydrochloride on ERG measured in rd mice.

Abstract

The invention concerns the use of a calcium channel blocker compound and/or cyclic GMP-dependent channels, namely diltiazem, for treating retinal pathologies, and more particularly retinal diseases caused by degeneration of visual receptors, in a human or animal.

Description

UTILISATION DU DILTIAZEM POUR LE TRAITEMENT DE PATHOLOGIESUSE OF DILTIAZEM FOR THE TREATMENT OF CONDITIONS
DE LA RETINEOF THE RETINA
La présente invention a pour objet l'utilisation de composés bloqueurs des canaux calciques et/ou de canaux activés par la guanosine monophosphate cyclique 3' 5' (cGMP) dans le cadre du traitement de pathologies de la rétine, et plus particulièrement des maladies de la rétine dues à une dégénérescence des photorécepteurs, chez l'homme ou l'animal, telles que la rétinite pigmentaire ou autres pathologies touchant semblablement les photorécepteurs, notamment la dégénérescence maculaire liée à l'âge.The present invention relates to the use of calcium channel blocking compounds and / or channels activated by cyclic guanosine monophosphate 3 '5' (cGMP) in the context of the treatment of pathologies of the retina, and more particularly of diseases of the retina due to a degeneration of photoreceptors, in humans or animals, such as retinitis pigmentosa or other pathologies similarly affecting photoreceptors, in particular macular degeneration linked to age.
La rétinite pigmentaire désigne un ensemble de ces maladies dégénératives des photorécepteurs (Berson, 1996) conduisant à la cécité. De nombreuses mutations affectant différentes protéines des bâtonnets, et susceptibles d'être à l'origine de cette maladie, ont été mises en évidence. Parmi ces mutations, on peut citer celles affectant les gènes de protéines impliquées dans la cascade de la phototransduction, telles que la rhodopsine, la transducine, la phosphodiestérase, rarrestine, ou de protéines structurales, telle que la périphérine. La souris rd (retinal degeneration) a été étudiée pendant plus de 70 ans comme modèle de la rétinite pigmentaire (Farber et coll., 1994) car le processus de dégénérescence rétinienne est similaire à celui observé dans la rétine pigmentaire, la mort des bâtonnets de la rétine étant suivie par une perte inexpliquée des cônes de la rétine. De plus, la mutation causale a été localisée dans le gène codant pour la sous-unité β de la cGMP-phosphodiestérase (PDE)Pigmentary retinitis is a group of these degenerative photoreceptor diseases (Berson, 1996) leading to blindness. Numerous mutations affecting different rod proteins, and likely to be at the origin of this disease, have been highlighted. Among these mutations, mention may be made of those affecting the genes of proteins involved in the phototransduction cascade, such as rhodopsin, transducin, phosphodiesterase, rarrestine, or of structural proteins, such as peripherin. The rd mouse (retinal degeneration) has been studied for more than 70 years as a model of pigmentary retinitis (Farber et al., 1994) because the process of retinal degeneration is similar to that observed in the pigment retina, the death of the rods of the retina being followed by an unexplained loss of cones in the retina. In addition, the causal mutation was located in the gene coding for the β subunit of cGMP-phosphodiesterase (PDE)
(Bowes et coll., 1990), comme dans certaines familles affectées par la maladie (Me Laughlin et coll., 1993).(Bowes et al., 1990), as in some families affected by the disease (Me Laughlin et al., 1993).
La PDE est activée durant la cascade de phototransduction par la chaîne α de la transducine, elle-même activée par la rhodopsine stimulée par la lumière. La PDE activée hydrolyse le cGMP, réduisant ainsi la concentration de cGMP et donc le nombre des canaux cGMP-dépendants ouverts, la conséquence finale étantPDE is activated during the phototransduction cascade by the α chain of transducin, itself activated by light-stimulated rhodopsin. Activated PDE hydrolyzes cGMP, thereby reducing the concentration of cGMP and therefore the number of open cGMP-dependent channels, the final consequence being
+ 2+ une diminution de la conductance aux cations tels que Na et Ca d'où une réduction du courant de dépolarisation des photorécepteurs dans l'obscurité. Chez la souris rd, Farber et Lolley (1974) ont montré qu'une augmentation anormale de la concentration de cGMP précède la dégénérescence des photorécepteurs. La toxicité du cGMP à forte concentration a ensuite été établie sur des photorécepteurs normaux (Lolley et Farber, 1977; Ulshafer et coll., 1980). Quelques approches thérapeutiques destinées à prévenir la perte de photorécepteurs sont actuellement en cours d'investigation sur les souris rd. Il a ainsi été décrit que la thérapie génique in vivo permet de retarder la mort des photorécepteurs pendant six semaines après injection sous-rétinienne d'un adénovirus recombinant à réplication défectueuse qui contient l'ADNc codant la+ 2+ a decrease in the conductance to cations such as Na and Ca hence a reduction in the depolarization current of the photoreceptors in the dark. In rd mice, Farber and Lolley (1974) have shown that an abnormal increase in the concentration of cGMP precedes the degeneration of photoreceptors. The toxicity of cGMP at high concentration was then established on normal photoreceptors (Lolley and Farber, 1977; Ulshafer et al., 1980). Some therapeutic approaches intended to prevent the loss of photoreceptors are currently being investigated in rd mice. It has thus been described that gene therapy in vivo makes it possible to delay the death of photoreceptors for six weeks after subretinal injection of a recombinant adenovirus with defective replication which contains the cDNA coding for
PDE murine (Bennett et coll., 1996). La transplantation de photorécepteurs (Gouras et coll., 1994, Silverman et coll., 1989) a été décrite comme permettant de préserver les photorécepteurs des cônes (Mohand-Said et coll., 1997). L'interprétation de cet effet en tant que mécanisme paracrine est en corrélation avec l'augmentation de la survie des photorécepteurs observée en coculture avec des photorécepteurs sains (Mohand-Said et coll., 1998) ou après application in vivo ou in vitro de facteurs trophiques tels que les facteurs de croissance fibroblastiques ou neuronaux (LaVail et coll., 1998).Murine PDE (Bennett et al., 1996). Transplantation of photoreceptors (Gouras et al., 1994, Silverman et al., 1989) has been described as making it possible to preserve the cone photoreceptors (Mohand-Said et al., 1997). The interpretation of this effect as a paracrine mechanism is correlated with the increase in photoreceptor survival observed in coculture with healthy photoreceptors (Mohand-Said et al., 1998) or after in vivo or in vitro application of factors trophics such as fibroblast or neuronal growth factors (LaVail et al., 1998).
Cependant, aucun traitement des maladies de la rétine dues à une dégénérescence des photorécepteurs, n'est actuellement disponible, mise à part la prescription de vitamine A pour la rétinite pigmentaire (Berson, 1996).However, no treatment for diseases of the retina due to degeneration of photoreceptors is currently available, apart from the prescription of vitamin A for retinitis pigmentosa (Berson, 1996).
La présente invention a précisément pour but de fournir des compositions pharmaceutiques utilisables dans le cadre du traitement de maladies de la rétine dues à une dégénérescence des photorécepteurs chez l'homme ou l'animal. En effet, la présente invention découle de la mise en évidence par lesThe object of the present invention is precisely to provide pharmaceutical compositions which can be used in the treatment of diseases of the retina due to degeneration of photoreceptors in humans or animals. Indeed, the present invention follows from the demonstration by the
Inventeurs du fait que les composés Moqueurs des canaux calciques et/ou cGMP- dépendants, tels que le chlorhydrate de diltiazem, permettent non seulement de ralentir la dégénérescence des bâtonnets et des cônes chez la souris rd, mais également de préserver la capacité des cellules rétiniennes à répondre aux stimuli lumineux.Inventors of the fact that the calcium channel and / or cGMP-dependent mocking compounds, such as diltiazem hydrochloride, make it possible not only to slow the degeneration of rods and cones in rd mice, but also to preserve the capacity of retinal cells to respond to light stimuli.
Ainsi, la présente invention a pour objet l'utilisation de composés bloqueurs des canaux calciques et/ou cGMP-dépendants pour la préparation d'un médicament destiné au traitement des pathologies liées à la dégénérescence des photorécepteurs de la rétine, et plus particulièrement de la rétinite pigmentaire, ou de pathologies touchant semblablement les photorécepteurs, telle que la dégénérescence maculaire liée à l'âge.Thus, the subject of the present invention is the use of calcium channel blocking and / or cGMP-dependent compounds for the preparation of a medicament intended for the treatment of pathologies linked to the degeneration of photoreceptors of the retina, and more particularly of the retinitis pigmentosa, or pathologies similarly affecting photoreceptors, such as age-related macular degeneration.
Par composés bloqueurs des canaux calciques et/ou cGMP-dépendants on entend tout composé capable de réduire la conductance ionique de ces canaux.By blocking compounds of calcium channels and / or cGMP-dependent is meant any compound capable of reducing the ionic conductance of these channels.
L'invention a plus particulièrement pour objet l'utilisation du diltiazem (D- cis-énantiomère), du L-cis-énantiomère, de leurs métabolites et de leurs sels pharmaceutiquement acceptables, pour la préparation d'un médicament destiné au traitement des pathologies liées à la dégénérescence des photorécepteurs de la rétine, notamment au traitement de la rétinite pigmentaire, ou de pathologies touchant semblablement les photorécepteurs, telle que la dégénérescence maculaire liée à l'âge.A more particular subject of the invention is the use of diltiazem (D-cis-enantiomer), L-cis-enantiomer, their metabolites and their pharmaceutically acceptable salts, for the preparation of a medicament intended for the treatment of pathologies linked to the degeneration of the photoreceptors of the retina, in particular to the treatment of retinitis pigmentosa, or pathologies similarly affecting photoreceptors, such as age-related macular degeneration.
L'invention concerne plus particulièrement l'utilisation susmentionnée du diltiazem de formuleThe invention relates more particularly to the aforementioned use of the diltiazem of formula
ainsi que ses sels d'addition aux acides pharmaceutiquement acceptables, notamment le malate ou le chlorhydrate de diltiazem.as well as its addition salts with pharmaceutically acceptable acids, in particular malate or diltiazem hydrochloride.
L'invention a également pour objet l'utilisation susmentionnée du L-cis énantiomère du diltiazem, ou le racemate du diltiazem, ainsi que leurs sels d'addition aux acides pharmaceutiquement acceptables, notamment le chlorhydrate de l'isomère cis(-) de formule suivante :The subject of the invention is also the above-mentioned use of the L-cis enantiomer of diltiazem, or the racemate of diltiazem, as well as their addition salts with pharmaceutically acceptable acids, in particular the hydrochloride of the cis (-) isomer of formula next :
L'invention concerne également l'utilisation susmentionnée de l'un des métabolites du diltiazem, ainsi que leurs sels d'addition aux acides pharmaceutiquement acceptables, répondant aux formules suivantes : The invention also relates to the abovementioned use of one of the metabolites of diltiazem, as well as their addition salts with pharmaceutically acceptable acids, corresponding to the following formulas:
Avantageusement, les composés bloqueurs des canaux calciques et/ou cGMP-dépendants susmentionnés sont utilisés pour la préparation de compositions pharmaceutiques se présentant sous une forme administrable par toute voie, notamment par voie orale, intramusculaire, intraveineuse, intraoculaire, ou sous forme de collyre.Advantageously, the above-mentioned calcium channel blocking and / or cGMP-dependent compounds are used for the preparation of pharmaceutical compositions which are in a form which can be administered by any route, in particular by oral, intramuscular, intravenous, intraocular, or in the form of eye drops.
De préférence les compositions pharmaceutiques de l'invention comprennent, sous forme unitaire, environ 0,1 à environ 100 mg d'un composé bloqueur des canaux calciques et/ou cGMP-dépendants, tel que défini ci-dessus, en association avec un véhicule pharmaceutiquement acceptable.Preferably, the pharmaceutical compositions of the invention comprise, in unit form, approximately 0.1 to approximately 100 mg of a calcium channel blocking compound and / or cGMP-dependent, as defined above, in combination with a vehicle pharmaceutically acceptable.
L'invention sera davantage illustrée à l'aide des figures 1 à 3 suivantes.The invention will be further illustrated with the aid of Figures 1 to 3 below.
La figure 1 illustre la survie des bâtonnets au 25° jour et au 36° jour postnatal chez les souris rd traitées par le chlorhydrate de diltiazem. Les injections de chlorhydrate de diltiazem ont débutés au 9° jour postnatal qui correspond à la période d'apparition des premiers signes de dégénérescence des bâtonnets. Les doses de chlorhydrate de diltiazem (2,5 mg/ml dans une solution physiologique) ont été augmentées graduellement à partir de 50μl par jour jusqu'à 100 μl deux fois par jour suivant la croissance de l'animal. Puisque les électrorétinogrammes (ERG) ont été mesurés avant le sacrifice des animaux, la dernière injection a été donnée 48 heures avant la mesure physiologique. Après fixation des rétines, les bâtonnets ont été marqués par des anticorps anti-rhodopsine (rho-4D2) (Hicks and Molday, 1986) et leur nombre a été estimé par stéréologie sur les rétines mises à plat en utilisant une procédure d'échantillonnage aléatoire (Mohand-Said et coll., 1998). Seules les rétines droites ont été prises en considération afin d'obtenir des résultats indépendants. Le traitement au chlorhydrate de diltiazem a augmenté de 86 % la survie des bâtonnets au 25° jour, et de 148 % au 36° jour chez les animaux traités par rapport aux souris rd de contrôle. Des injections répétées de solution physiologique seule n'ont pas affecté de façon significative la survie des bâtonnets (7416 + 1291, s.e.m., n = 5) par rapport aux animaux non traités (7648 + 774, s.e.m., n = 4). Ces observations montrent donc que le traitement des souris rd par le chlorhydrate de diltiazem induit une survie des bâtonnets.FIG. 1 illustrates the survival of the rods at the 25th day and at the 36th postnatal day in rd mice treated with diltiazem hydrochloride. The injections of diltiazem hydrochloride started on the 9th postnatal day which corresponds to the period of appearance of the first signs of degeneration of the rods. The doses of diltiazem hydrochloride (2.5 mg / ml in physiological solution) were gradually increased from 50 μl per day to 100 μl twice a day following the growth of the animal. Since the electroretinograms (ERG) were measured before the animals were sacrificed, the last injection was given 48 hours before the physiological measurement. After retinal fixation, the rods were labeled with anti-rhodopsin antibodies (rho-4D2) (Hicks and Molday, 1986) and their number was estimated by stereology on the retinas laid flat using a random sampling procedure. (Mohand-Said et al., 1998). Only the right retinas were taken into account in order to obtain independent results. Treatment with diltiazem hydrochloride increased the survival of the rods at day 25 by 86%, and by 148% at day 36 in the treated animals compared to the control rd mice. Repeated injections of physiological solution alone did not significantly affect the survival of the rods (7416 + 1291, wk, n = 5) compared to untreated animals (7648 + 774, wk, n = 4). These observations therefore show that the treatment of rd mice with diltiazem hydrochloride induces rod survival.
La figure 2 illustre la survie des cônes au 25° jour et au 36° jour postnatal chez les souris rd traitées par le chlorhydrate de diltiazem. Le nombre de cônes a été estimé indirectement après coloration des noyaux avec le DAPI (4',6-diamino- 2-phénylindole) sur des coupes de rétines. Pour obtenir ce nombre, le nombre de bâtonnets immunomarqués a été soustrait du nombre de noyaux de photorécepteurs marqués au DAPI. Le traitement au chlorhydrate de diltiazem a augmenté de 109 % la survie des cônes au 25° jour, et de 144 % au 36° jour chez les animaux traités par rapport aux souris rd de contrôle.FIG. 2 illustrates the survival of the cones on the 25th day and on the 36th postnatal day in rd mice treated with diltiazem hydrochloride. The number of cones was estimated indirectly after staining of the nuclei with DAPI (4 ', 6-diamino-2-phenylindole) on sections of retina. To obtain this number, the number of immunostained rods was subtracted from the number of photoreceptor nuclei labeled with DAPI. Treatment with diltiazem hydrochloride increased the survival of the cones at the 25th day by 109% and by 144% at the 36th day in the treated animals compared to the control rd mice.
La figure 3 illustre le fait que la survie des photorécepteurs est accompagnée d'améliorations physiologiques chez les souris rd traitées par le chlorhydrate de diltiazem. Cette démonstration a été effectuée par mesure des ERG chez les souris rd traitées et les souris rd de contrôle. Chez des souris rd non traitées, les amplitudes des ondes a et b des ERG diminuent régulièrement à partir du 12° jour postnatal jusqu'à extinction au 24° jour postnatal. En revanche, tous les animaux traités présentent des signaux ERG AU 25° jour postnatal (n = 7). Au 36° jour postnatal, 4 animaux traités sur 10 présentent des signaux ERG pouvant être mesurés dans les deux yeux. Ces observations montrent que le traitement par le chlorhydrate de diltiazem non seulement permet aux bâtonnets de survivre mais protège aussi les fonctions visuelles de la rétine. FIG. 3 illustrates the fact that the survival of photoreceptors is accompanied by physiological improvements in rd mice treated with diltiazem hydrochloride. This demonstration was carried out by measurement of ERG in the rd treated mice and the rd control mice. In untreated rd mice, the amplitudes of the a and b waves of the ERGs decrease regularly from the 12th postnatal day until extinction on the 24th postnatal day. On the other hand, all the treated animals show ERG signals AU 25 ° postnatal day (n = 7). At 36 ° postnatal day, 4 animals out of 10 treated show ERG signals which can be measured in both eyes. These observations show that treatment with diltiazem hydrochloride not only allows the rods to survive but also protects the visual functions of the retina.
Légende des figures :Legend of figures:
- Figure 1 : estimation du nombre de bâtonnets sur des rétines entières de souris rd de 25 jours et de 36 jours traitées par le chlorhydrate de diltiazem, par rapport aux souris rd de contrôle non traitées.- Figure 1: estimate of the number of rods on whole retinas of rd mice of 25 days and 36 days treated with diltiazem hydrochloride, compared to untreated control rd mice.
- Figure 2 : estimation du nombre de cônes et de bâtonnets sur des coupes de rétines de souris rd de 25 jours et de 36 jours traitées par le chlorhydrate de diltiazem, par rapport aux souris rd de contrôle non traitées.- Figure 2: estimation of the number of cones and rods on sections of retina of rd mice of 25 days and 36 days treated with diltiazem hydrochloride, compared to untreated control rd mice.
- Figure 3 : effet du chlorhydrate de diltiazem sur l'ERG mesuré chez les souris rd. A) représentation des enregistrements d'ERG mesurés chez les souris rd traitées et de contrôle au 25° et au 36° jour postnatal. B) effet du chlorhydrate de diltiazem sur l'onde b de l'ERG ; la courbe correspondant à la mesure de l'amplitude de l'onde b en fonction du temps chez les souris rd de contrôle est représentée à l'aide de cercles creux, tandis que celle correspondant à cette même mesure chez les souris rd traitées par le chlorhydrate de diltiazem est représentée à l'aide de carrés noirs. - Figure 3: effect of diltiazem hydrochloride on ERG measured in rd mice. A) representation of ERG recordings measured in rd treated and control mice at 25 ° and 36 ° postnatal day. B) effect of diltiazem hydrochloride on the b wave of the ERG; the curve corresponding to the measurement of the amplitude of the b wave as a function of time in the rd control mice is represented using hollow circles, while that corresponding to this same measurement in the rd mice treated with the diltiazem hydrochloride is shown using black squares.
REFERENCES BIBLIOGRAPHIQUESBIBLIOGRAPHICAL REFERENCES
Bennett J., Tanabe T., Sun D., Zeng Y., Kjeldbye H., Gouras P., Maguire A.M. ; "Photoreceptor cell rescue in retinal degeneration (rd) mice by in vitro gène therapy", Nat. Med. (1996) 2 : 649-654.Bennett J., Tanabe T., Sun D., Zeng Y., Kjeldbye H., Gouras P., Maguire A.M.; "Photoreceptor cell rescue in retinal degeneration (rd) mice by in vitro gene therapy", Nat. Med. (1996) 2: 649-654.
Berson E.L. ; "Retinitis pigmentosa : Unfolding its mystery", Proc. Natl. Acad. Sci. (1996) 93 : 4526-4528.Berson E.L.; "Retinitis pigmentosa: Unfolding its mystery", Proc. Natl. Acad. Sci. (1996) 93: 4526-4528.
Bowes C, Li T., Danciger M., Baxter L.C., Applebury M.L., Farber D. ; "Retinal degeneration in the rd mouse is caused by a defect in the β-subunit of rod cGMP- phosphodiesterase" ; Nature (1990) 347 : 677-680.Bowes C, Li T., Danciger M., Baxter L.C., Applebury M.L., Farber D.; "Retinal degeneration in the rd mouse is caused by a defect in the β-subunit of rod cGMP- phosphodiesterase"; Nature (1990) 347: 677-680.
Farber D.B. and Lolley R.N. ; Cyclic guanoside monophosphate : Elévation in degenerating photoreceptor cells of the C3H mouse retina. Science (1974) 186 : 449-451.Farber D.B. and Lolley R.N.; Cyclic guanoside monophosphate: Elevation in degenerating photoreceptor cells of the C3H mouse retina. Science (1974) 186: 449-451.
Farber D.B., Flannery J.G., Bowes-Rickman C. ; "The rd mouse story : seventy years of research on an animal model of inherited retinal degeneration", Prog. Ret.Farber D.B., Flannery J.G., Bowes-Rickman C.; "The rd mouse story: seventy years of research on an animal model of inherited retinal degeneration", Prog. Ret.
Eye Res. (1994) 31-62.Eye Res. (1994) 31-62.
Gouras P., Du J., Kjeldbye H., Yamamoto S., Zack D.J. ; "Long-term photoreceptor transplants in dystrophic and normal mouse retina", Invest. Ophthalmol. Vis. Sci. (1994) 35 : 3145-53.Gouras P., Du J., Kjeldbye H., Yamamoto S., Zack D.J.; "Long-term photoreceptor transplants in dystrophic and normal mouse retina", Invest. Ophthalmol. Screw. Sci. (1994) 35: 3145-53.
Hicks D. and Molday R.S. ; Differential immunogold-dextran labeling of bovine and frog rod and cônes cells using monoclonal antibodies against bovine rhodopsin. Expérimental Eye Research (1986) 42 : 55-71.Hicks D. and Molday R.S.; Differential immunogold-dextran labeling of bovine and frog rod and cones cells using monoclonal antibodies against bovine rhodopsin. Experimental Eye Research (1986) 42: 55-71.
Lavail M. M., Yasumura D., Matthes M. T., Lau-Villacorta C, Unoki K., Sung C. H., Steinberg R. H. Protection of mouse photoreceptors by survival factors in retinal degenerations. Invest. Ophthal. Vis. Sci. (1998) 39 : 592-602.Lavail M. M., Yasumura D., Matthes M. T., Lau-Villacorta C, Unoki K., Sung C. H., Steinberg R. H. Protection of mouse photoreceptors by survival factors in retinal degenerations. Invest. Ophthal. Screw. Sci. (1998) 39: 592-602.
Lolley R.N., Farber D.B., Rayborn M.E. and Hollyfield J.G. ; Cyclic GMP accumulation causes degeneration of photoreceptor cells : simulation of an inherited disease. Science (1977) 196 : 664-666. McLaughlin M.E., Sandberg M.A., Berson E.L., Drya T.P. ; "Récessive mutations in the gène encoding the b-subunit of rod phosphodiesterase in patients with retinitis pigmentosa", Nature genetics (1993) 4 : 130-133.Lolley RN, Farber DB, Rayborn ME and Hollyfield JG; Cyclic GMP accumulation causes degeneration of photoreceptor cells: simulation of an inherited disease. Science (1977) 196: 664-666. McLaughlin ME, Sandberg MA, Berson EL, Drya TP; "Recessive mutations in the gene encoding the b-subunit of rod phosphodiesterase in patients with retinitis pigmentosa", Nature genetics (1993) 4: 130-133.
Mohand-Said S., Hicks D., Simonutti M., Tran-Minh D., Deudon-Combe A.,Mohand-Said S., Hicks D., Simonutti M., Tran-Minh D., Deudon-Combe A.,
Dreyfus H., Silvermann M.S., Mosinger Ogilvie J., Tenkova T., Sahel J. ; "Photoreceptor transplants increase host cône survival in the retinal degeneration (rd) mouse", Ophth. Res. (1997) 29 : 290-297.Dreyfus H., Silvermann M.S., Mosinger Ogilvie J., Tenkova T., Sahel J.; "Photoreceptor transplants increase host cone survival in the retinal degeneration (rd) mouse", Ophth. Res. (1997) 29: 290-297.
Mohand-Said S., Deudon-Combe A., Hicks D., Simonutti M., Forster V., FintzMohand-Said S., Deudon-Combe A., Hicks D., Simonutti M., Forster V., Fintz
A.C., Léveillard T., Dreyfus H. and Sahel J. ; Normal retina releases a diffusible factor stimulating cône survival in the retinal degeneration mouse. Proceeding of the National Academy of Sciences (USA) (1998) 95 : 8357-8362.A.C., Léveillard T., Dreyfus H. and Sahel J.; Normal retina releases a diffusible factor stimulating cone survival in the retinal degeneration mouse. Proceeding of the National Academy of Sciences (USA) (1998) 95: 8357-8362.
Ulshafer R.J., Garcia C.A. and Hollyfield J.G. ; Sensitivity of photoreceptors to elevated levels of cGMP in the hurnan retina. Investigative Ophthalmology and Visual Science (1980) vol. 19, n° 10 : 1236-1241. Ulshafer R.J., Garcia C.A. and Hollyfield J.G.; Sensitivity of photoreceptors to elevated levels of cGMP in the hurnan retina. Investigative Ophthalmology and Visual Science (1980) vol. 19, no.10: 1236-1241.

Claims

REVENDICATIONS
1. Utilisation du diltiazem (D-cis-énantiomère), du L-cis-énantiomère, de leurs métabolites et de leurs sels pharmaceutiquement acceptables, pour la préparation d'un médicament destiné au traitement des pathologies liées à la dégénérescence des photorécepteurs de la rétine.1. Use of diltiazem (D-cis-enantiomer), L-cis-enantiomer, their metabolites and their pharmaceutically acceptable salts, for the preparation of a medicament intended for the treatment of pathologies linked to the degeneration of photoreceptors of the retina.
2. Utilisation du diltiazem (D-cis-énantiomère), du L-cis-énantiomère, de leurs métabolites et de leurs sels pharmaceutiquement acceptables, pour la préparation d'un médicament destiné au traitement de la rétinite pigmentaire, ou de pathologies touchant semblablement les photorécepteurs, telle que la dégénérescence maculaire liée à l'âge.2. Use of diltiazem (D-cis-enantiomer), L-cis-enantiomer, their metabolites and their pharmaceutically acceptable salts, for the preparation of a medicament intended for the treatment of pigmentary retinitis, or pathologies similarly affecting photoreceptors, such as age-related macular degeneration.
3. Utilisation selon la revendication 1 ou 2, du diltiazem de formule3. Use according to claim 1 or 2, of the diltiazem of formula
ainsi que de ses sels d'addition aux acides pharmaceutiquement acceptables, notamment le malate ou le chlorhydrate de diltiazem.as well as its addition salts with pharmaceutically acceptable acids, in particular malate or diltiazem hydrochloride.
4. Utilisation selon la revendication 1 ou 2, du L-cis énantiomère, ou du racemate, ainsi que leurs sels d'addition aux acides pharmaceutiquement acceptables. 4. Use according to claim 1 or 2, L-cis enantiomer, or racemate, as well as their addition salts with pharmaceutically acceptable acids.
5. Utilisation selon la revendication 1 ou 2, de l'un des métabolites du diltiazem et de l'autre énantiomère, ainsi que leurs sels d'addition aux acides pharmaceutiquement acceptables, répondant à l'une des formules suivantes :5. Use according to claim 1 or 2, of one of the metabolites of diltiazem and of the other enantiomer, as well as their addition salts with pharmaceutically acceptable acids, corresponding to one of the following formulas:
6. Utilisation selon l'une des revendications 1 à 5, pour la préparation d'un médicament se présentant sous une forme administrable par voie orale, intramusculaire, intraveineuse, intraoculaire, ou sous forme de collyre. 6. Use according to one of claims 1 to 5, for the preparation of a medicament which is in a form which can be administered orally, intramuscularly, intravenously, intraocularly, or in the form of eye drops.
EP99946279A 1998-10-02 1999-10-01 Use of diltiazem for treating retinal pathologies Expired - Lifetime EP1117408B9 (en)

Applications Claiming Priority (3)

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FR9812364A FR2784030B1 (en) 1998-10-02 1998-10-02 USE OF CALCIUM AND / OR CGMP-DEPENDENT CHANNEL BLOCKERS FOR THE TREATMENT OF RETINE CONDITIONS
FR9812364 1998-10-02
PCT/FR1999/002346 WO2000020006A1 (en) 1998-10-02 1999-10-01 Use of diltiazem for treating retinal pathologies

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Cited By (4)

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WO2013010854A2 (en) 2011-07-15 2013-01-24 Solvay Specialty Polymers Italy S.P.A. Fluoropolymer composition for multilayer assemblies
WO2019224489A1 (en) 2018-05-23 2019-11-28 Universite Claude Bernard Lyon 1 Diltiazem for use in the treatment of microbial infections
FR3106055A1 (en) 2020-01-13 2021-07-16 Centre National De La Recherche Scientifique (Cnrs) COMBINATION OF DILTIAZEM AND OTHER ANTIVIRAL AGENTS
WO2021181044A1 (en) 2020-03-10 2021-09-16 Universite Claude Bernard Lyon 1 Composition comprising diltiazem for treating a viral infection caused by sars-cov-2 viruses

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FR2823221B1 (en) 2001-04-06 2004-04-02 Univ Pasteur SEQUENCES ASSOCIATED WITH RETINAL DEGENERATION AND APPLICATIONS
DE60212729T2 (en) 2002-10-01 2007-06-28 Paola Ammannati ENZYMATIC TREATMENT OF RETINITIS PIGMENTOSA AND PHARMACEUTICAL COMPOSITION FOR THIS IN THE FORM OF A KIT
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US20110021974A1 (en) * 2010-10-05 2011-01-27 Shantha Totada R Retinitis pigmentosa treatment and prophalaxis
RU2716977C2 (en) 2013-07-31 2020-03-17 Новартис Аг Novel selective vectors and methods for selecting eukaryotic host cells
JP6954582B2 (en) * 2016-12-22 2021-10-27 国立大学法人滋賀医科大学 Pharmaceutical composition
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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013010854A2 (en) 2011-07-15 2013-01-24 Solvay Specialty Polymers Italy S.P.A. Fluoropolymer composition for multilayer assemblies
WO2019224489A1 (en) 2018-05-23 2019-11-28 Universite Claude Bernard Lyon 1 Diltiazem for use in the treatment of microbial infections
FR3106055A1 (en) 2020-01-13 2021-07-16 Centre National De La Recherche Scientifique (Cnrs) COMBINATION OF DILTIAZEM AND OTHER ANTIVIRAL AGENTS
WO2021144528A1 (en) 2020-01-13 2021-07-22 Universite Claude Bernard Lyon 1 Combination of diltiazem and viral polymerase inhibitors
WO2021181044A1 (en) 2020-03-10 2021-09-16 Universite Claude Bernard Lyon 1 Composition comprising diltiazem for treating a viral infection caused by sars-cov-2 viruses
FR3108033A1 (en) 2020-03-10 2021-09-17 Universite Claude Bernard Lyon 1 ANTIVIRAL COMPOUNDS AND THEIR COMBINATIONS FOR TREATING VIRAL INFECTION WITH SARS-CoV-2

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JP2002526411A (en) 2002-08-20
ATE269085T1 (en) 2004-07-15
FR2784030B1 (en) 2002-12-20
DE69918145D1 (en) 2004-07-22
EP1117408B1 (en) 2004-06-16
WO2000020006A1 (en) 2000-04-13
ES2219060T3 (en) 2004-11-16
DE69918145T2 (en) 2005-08-11
FR2784030A1 (en) 2000-04-07

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